To distinguish between live and dead cells, the LIVE/DEAD Viability/Cytotoxicity Kit for mammalian cells (Life Technologies GmbH) was performed according to the manufacturer's instructions. At first, cell nuclei were counterstained with 2 µg/ml cell-permeant nucleic acid stain bisBenzimide H 33342 trihydrochloride (Hoechst 33342; Sigma-Aldrich Chemie GmbH) for 30 min at 37°C. In the following, cells were incubated with 2 nM calcein AM and 4 nM ethidium homodimer-1 for 30 min at 37°C. After washing twice with PBS, cells were examined using the inverted microscope Axio Observer (Carl Zeiss Microscopy GmbH).
Bisbenzimide h 33342 trihydrochloride hoechst 33342
Manufactured by Merck Group
Sourced in United States
Bisbenzimide H 33342 trihydrochloride (Hoechst 33342) is a fluorescent dye that binds to adenine-thymine (A-T) rich regions in DNA. It is a cell-permeable nuclear and chromosome counterstain that is frequently used in fluorescence microscopy and flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Live dead viability cytotoxicity kit for mammalian cells, by Thermo Fisher Scientific (1 mentions)
Axio observer, by Zeiss (1 mentions)
Fluorescein isothiocyanate isomer, by Merck Group (1 mentions)
Pei 25k, by Merck Group (1 mentions)
Pei 750k, by Merck Group (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Apaf 1, by Cell Signaling Technology (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Parp 1, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Bcl xl, by Santa Cruz Biotechnology (1 mentions)
Monodansylcadaverine, by Merck Group (1 mentions)
Zinc pyrithione, by Merck Group (1 mentions)
Click it edu kit, by Thermo Fisher Scientific (1 mentions)
Newport green dcf diacetate, by Thermo Fisher Scientific (1 mentions)
Cyclosporin a, by Merck Group (1 mentions)
Acridine orange, by Merck Group (1 mentions)
8 protocols using bisbenzimide h 33342 trihydrochloride hoechst 33342
1
Live/Dead Cell Viability Assay
2
Functionalized Nanomaterials Synthesis
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3-Mercaptopropyltrimethoxysilane (MPMS), 3-aminopropyltrimethoxysilane (APS), fluorescein isothiocyanate isomer (FITC), rhodamine B, bisbenzimide H 33342 trihydrochloride (Hoechst 33342) and branched polyethyleneimine solution [MW = 1.3k (PEI1.3k), 2.0k (PEI2.0k), 25k (PEI25k), and 750k (PEI750k)] were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Ammonium hydroxide (NH4OH, 28%) and ethanol were purchased from Kishida Chemical (Osaka, Japan). All other reagents and solvents were of analytical grade.
3
Investigating Ovarian Cancer Cell Lines
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Human immortalized ovarian surface epithelial cells (IOSE 364) and human ovarian carcinoma cells (OVCAR-3) were donated by Dr. Auersperg (University of British Columbia, Vancouver, BC, Canada) and Dr. Jiang (Thomas Jefferson University, Philadelphia, PA, USA), respectively. Both cells were grown in RPMI 1640 medium containing 10% fetal bovine serum.
Reagents: TF3 monomer (purity of 92.4%) was prepared by a previously established method [27 (link)]. BisBenzimide H 33342 trihydrochloride (Hoechst 33342) were purchased from Sigma (St. Louis, MO, USA). Propidium iodide (PI) was purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against phospho-Chk2 (Thr68), Chk2, p53, Bad, Bcl-xL, PARP-1 and GAPDH were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Antibodies against Bax, Bcl-2, Mcl-1, Apaf-1, Fas, DR5, FADD, p27, CDK4, Cyclin D1, p-Rb and Rb were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).
Reagents: TF3 monomer (purity of 92.4%) was prepared by a previously established method [27 (link)]. BisBenzimide H 33342 trihydrochloride (Hoechst 33342) were purchased from Sigma (St. Louis, MO, USA). Propidium iodide (PI) was purchased from Invitrogen (Carlsbad, CA, USA). Antibodies against phospho-Chk2 (Thr68), Chk2, p53, Bad, Bcl-xL, PARP-1 and GAPDH were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Antibodies against Bax, Bcl-2, Mcl-1, Apaf-1, Fas, DR5, FADD, p27, CDK4, Cyclin D1, p-Rb and Rb were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).
4
Apoptosis and Necrosis Assay Protocol
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JC-1, Newport Green™ DCF diacetate, DQ-Green BSA and Click-iT EdU Kit were acquired from Molecular Probes, Inc. (Eugene, OR, USA). Zinc pyrithione, acridine orange, cyclosporin A, monodansylcadaverine (MDC), N-acetyl cysteine (NAC), 3-methyladenine (3-MA), chloroquine (CQ), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS), horseradish peroxidase, Triton-X, dithiotreitol (DTT), bisBenzimide H 33342 trihydrochloride (Hoechst 33342) and 4’,6-Diamidino-2-Phenylindole (DAPI) were obtained from Sigma-Aldrich (St. Louis, MO, USA). WST-1 was purchased from Roche Diagnostics (Manheim, Germany). Caspase-3 inhibitor z-devd-fmk was from ICN Biomedicals Inc. (Irvine, CA, USA). Necrostatin-1 was from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Primary and secondary antibodies were from Cell Signaling Technology (Danvers, MA, USA). All other chemicals were of the highest analytical grade.
5
Extracellular LRP6 Domain Detection
6
Fluorescent Labeling for Cell Tracking
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Medium-199, CellTracker TM Red, and 2 0 ,7 0 -dichlorodihydrofluorescein diacetate (Carboxy-DCFDA) were purchased from Invitrogen (Invitrogen, Carlsbad, CA, USA). Luteinizing hormone (LH), follicle-stimulating hormone (FSH), epidermal growth factor (EGF), bovine serum albumin (BSA), and bisbenzimide H 33342 trihydrochloride (Hoechst 33342) were obtained from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA).
7
Multimodal Imaging of Cell Morphology
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Cells were fixed with 4% paraformaldehyde
(Sigma-Aldrich) at 4 °C, permeabilized with 0.1% triton X-100
(Sigma-Aldrich) at room temperature, and blocked with bovine serum
albumin 2% (Sigma-Aldrich) for 45 min at room temperature. Then, cells
were incubated with DAPI (Molecular probes) for nucleus labeling and
actin-green (molecular probes) for cytoskeleton labeling for 20 min
at room temperature. To visualize the living cells during the NP flow
studies, they were labeled with bisBenzimide H 33342 trihydrochloride
(Hoechst 33342, Sigma-Aldrich). SEM images (FSEM Auriga, Carl Zeiss)
were taken after a thin layer of gold was sputter-coated.
(Sigma-Aldrich) at 4 °C, permeabilized with 0.1% triton X-100
(Sigma-Aldrich) at room temperature, and blocked with bovine serum
albumin 2% (Sigma-Aldrich) for 45 min at room temperature. Then, cells
were incubated with DAPI (Molecular probes) for nucleus labeling and
actin-green (molecular probes) for cytoskeleton labeling for 20 min
at room temperature. To visualize the living cells during the NP flow
studies, they were labeled with bisBenzimide H 33342 trihydrochloride
(Hoechst 33342, Sigma-Aldrich). SEM images (FSEM Auriga, Carl Zeiss)
were taken after a thin layer of gold was sputter-coated.
8
Purification and Analysis of Apoptosis Regulators
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